Endothelial NO synthase (eNOS, NOS-III) belongs to a group of three isoenzymes which produce nitric oxide (nitrogen monoxide, NO) by oxidation of arginine. Endothelially released NO is of central importance in a number of key cardiovascular mechanisms. It has a vasodilating effect and inhibits the aggregation of platelets, the adhesion of leukocytes to the endothelium and the proliferation of intimal smooth muscle cells.
Endothelial NO synthase is subject to physiological and pathophysiological regulation both at the transcriptional and at the post-transcriptional level. Enzyme already present in the endothelium may undergo calcium-dependent and calcium-independent activation through phosphorylation of specific amino acids, but also by direct interactions with specific proteins. Stimulators of this, usually transient, NO release are extracellular arginine, 17β-estrogen and the mechanical stimulus exerted on the luminal surface of the endothelium by the blood flow (shear stress). The latter additionally leads to regulation of eNOS at the transcriptional level. Thus, for example, Sessa et al. (Circ. Research 74 (1994) 349) were able to obtain a marked increase in eNOS by means of exercise training and the increase in shear stress associated therewith.
Whether regulation at the post-transcriptional level is relevant in vivo, has not been unambiguously proven. Thus, for example, administration of a high arginine dose is followed by only a transient improvement in the endothelium-dependent vasorelaxation in patients with coronary heart disease.
On the other hand, the significance of the upregulation of the eNOS protein is scientifically accepted. Thus, there are findings which show that the protective properties of the HMG-CoA reductase inhibitor simvastatin can be attributed, besides to the lipid lowering, also in part to an increase in eNOS expression in vivo (Endres et al., Proc. Natl. Acad. Sci. USA 95 (1998) 8880). It is additionally known that single point mutations in the 5′-flanking region of the eNOS gene (“eNOS promoter”), and the reduction in the rate of eNOS gene transcription associated therewith, in the Japanese population is associated with an increase in the risk of coronary spasms (Nakayama et al., Circulation 99 (1999) 2864).
The current assumption therefore is that the transcriptional and post-transcriptional mechanisms of eNOS regulation are seriously disturbed in a large number of disorders, especially in cardiovascular disorders. Even in very early stages of a wide variety of cardiovascular disorders it is possible for a dysfunction of this type in the endothelium lining the blood vessels to lead to a deficiency of bioactive NO, which is manifested as the disorder progresses in the form of measurable pathophysiological and morphological changes. Thus, critical steps in early atherogenesis are speeded up by a decrease in endothelial NO release, such as, for example, the oxidation of low density lipoproteins, the recruitment and deposition of monocytes in the intima of vessels, and the proliferation of intimal cells. A consequence of atherogenesis is the formation of plaques on the inside of the blood vessels, which may in turn lead, through a diminution in the shear stress, to a further decrease in endothelial NO release and a further deterioration in the pathology. Since endothelial NO is also a vasodilator, a decrease thereof frequently also leads to hypertension which may, as an independent risk factor, cause further organ damage.
The aim of a therapeutic approach to the treatment of these disorders must accordingly be to interrupt this chain of events by increasing the endothelial NO expression. Gene transfer experiments which lead in vitro to overexpression of NO synthase in previously damaged vessels are in fact able to counteract the described processes and are thus evidence of the correctness of this approach (Varenne et al., Hum. Gene Ther. 11 (2000) 1329).
Some low molecular weight compounds which, in cell cultures, may lead to a direct effect on eNOS transcription and expression are disclosed in the literature. For the statins, as has already been mentioned, it has been possible to show such an increase in eNOS in vivo as a side effect. In view of the known range of side effects of this class of substances, however, it is unclear how far use of this effect can be made in a toxicologically unproblematic dose. Liao et al. claim in WO 99/47153 and WO 00/03746 the use of rhoGTPase inhibitors and agents which influence the organization of the actin cytoskeleton for increasing eNOS in endothelial cells and for the therapy of various disorders such as, for example, strokes or pulmonary hypertension without, however, indicating a specific way of achieving this. Certain amide derivatives which upregulate the expression of endothelial NO synthase, in particular N-cycloalkyl amides in which the cycloalkyl ring is fused to a benzene ring or a heteroaromatic ring, have been described in WO 02/064146, WO 02/064545, WO 02/064546, WO 02/064565, WO 2004/014369, WO 2004/014372 and WO 2004/014842. Certain triaza- and tetraaza-anthracenedione derivatives which upregulate the expression of endothelial NO synthase have been described in WO 2004/094425. There still exists a need for further compounds which upregulate the expression of endothelial NO synthase and have a favorable property profile and are useful as pharmaceuticals for the treatment of various diseases such as atherosclerosis, coronary artery disease or cardiac insufficiency, for example. Surprisingly it has now been found that the compounds of the formula I are modulators of the transcription of endothelial NO synthase and in particular stimulate, or upregulate, the expression of eNOS, and are useful for the treatment of various diseases such as the mentioned cardiovascular disorders.
Certain compounds which are encompassed by the formula I have already been described. In JP 2004-262890 compounds are described which inhibit the synthesis of 20-hydroxyeicosatetraenoic acid (=20-HETE) from arachidonic acid and which contain a substituted central benzene ring or pyridine ring. The central ring carries a heterocyclic group, which in the case of a central pyridine ring is present in the 5-position, and a further group of broad structural variety, which in the case of a central pyridine ring is present in the 2-position. Said further group can, among others, be an unsubstituted imidazolin-2-on-1-yl group, which is assumed to mean a 2-oxoimidazolin-1-yl group, or an unsubstituted 2-oxopyrrolidin-1-yl group which are linked to the central ring via an alkyloxy group. Compounds of the formula I, in which R1 and R2 together with the N—CO group which carries them form a group of the formula
i.e. compounds in which the group —N(R2)—CO—R1 is a phthalimido group which group is also designated as 1,3-dioxoisoindol-2-yl group or 1,3-dioxo-1,3-dihydroisoindol-2-yl group or 1,3-dioxo-2,3-dihydro-1H-isoindol-2-yl group, are described in a number of documents including DE 1913471, GB 1341375, U.S. Pat. No. 5,190,589, U.S. Pat. No. 5,547,972 and Lipinski et al., J. Med. Chem. 29 (1986) 2154. Such phthalimido compounds commonly are intermediates in the synthesis of the respective compounds containing an amino group —NH2 using the Gabriel reaction, or are protected forms of such amino compounds.
In other documents certain compounds comprised by the formula I are described in which R1 and R2, together with the N—CO group which carries them, do not form a ring. For example, In U.S. Pat. No. 5,292,732 1-(2-acylaminoethyl)pyrroles are described which are intermediates in the synthesis of monoamine oxidase inhibiting pyrrolopyrazines. In JP 44-29656 4-(ω-acylaminoalkyl)isoxazoles are described which exhibit analgesic, antitussive, antipyretic and antiinflammatory activity. In U.S. Pat. No. 3,655,684 3-(2-acylaminoethyl)-5-phenyl-1,2,4-oxadiazoles are described which exhibit anticonvulsant activity. Certain 3-cyanopyridines which carry an oxaalkylamino, aminoalkylamino, oxaalkyloxy, acyloxyalkylamino, acylaminoalkylamino or acyloxyalkyloxy group in the 2-position and a pyridin-4-yl group or an alkyloxy-substituted phenyl group in the 5-position and which exhibit cardiotonic activity, including the specific compounds of the formula I in which A is the group NH—CH2—CH2 the nitrogen atom of which is attached to the group Het, Het is a pyridinyl group whose 2-position carries the group A and which is substituted by cyano in the 3-position, X is a direct bond, R3 is pyridin-4-yl which is attached to the 5-position of the pyridinyl group representing Het, R2 is hydrogen and R1 is either methyl or ethyl or pyridin-3-yl, are described in Hagen et al., Pharmazie 45 (1990) 343 and EP 200024. A stimulating effect of these known compounds of the formula I on the transcription or the expression of eNOS and their use in the treatment of diseases which is based on such effect, has not yet been described.